Efficient In Vitro Shoot-regeneration Systems in Cassava (Manihot esculenta Crantz)

Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA₃, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava.     

Immunization of cattle with nymphal Hyalomma anatolicum anatolicum extracts: effects on tick biology

Antigens derived from partially engorged nymphs of Hyalomma anatolicum anatolicum were used in immunizing crossbred (Bos indicus×Bos taurus) cattle against larval, nymphal and adult H. a. anatolicum and H. dromedarii. The cattle were either infected with Theileria annulata at low parasitaemia or were uninfected. Whole nymphal extract (WNE), nymphal membrane antigens (NMA) and nymphal soluble antigens (NSA) were used for immunization. The group immunized with WNE showed significant and better rejection of H. a. anatolicum ticks as compared to calves immunized with either NMA or NSA. The moulting rates of both engorged larvae and nymphs remained unaffected. Nymphs which engorged on the immunized calves were fully susceptible to infection by T. annulata as indicated by the intensity and abundance of Theileria infections in the resulting adult ticks from immunized and unimmunized Theileria infected cattle. These ticks also transmitted fatal theileriosis to susceptible calves.     

Folacin and cyanocobalamin in relation to naturalTrypanosoma evansi Infection in buffaloes

Summary The haematological values of haemoglobin (Hb), packed cell volume (PCV) and vitamin levels of folacin (folic acid), cyanocobalamin (vitamin B₁₂) of buffaloes, in a surra endemic area of Eastern Haryana, India, were determined. Surra-positive buffaloes had significantly low levels of Hb, PCV, folic acid and vitamin B₁₂. These low levels of folic acid and vitamin B₁₂ may have enhanced the clinical signs of chronic trypanosomosis, caused byTrypanosoma evansi.

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Folacina Y Cianocobalamina En Relacion Con La Infeccion Natural De Bufalos ConTrypanosoma Evansi

Resumen Se determinaron los valores hematológicos de hemoglobina (Hb), volumen corpuscular celular (PCV) y los niveles vitamínicos de ácido fólico y cianocobalamina (vitamina B₁₂) de búfalos, en un área endémica de surra en Haryana Oriental, India. Los búfalos positivos a surra tuvieron niveles bajos de Hb, PCV, ácido fólico y vitamina B₁₂. Los niveles bajos de ácido fólico y cianocobalamina, pudieron haber exacerbado los sintomas clinicos de tripanosomiasis crónica, causada porTrypanosoma evansi.

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Folacine Et Cyanocobalamine En Relation Avec La Trypanosomose ATrypanosoma Evansi Chez Des Buffles

Résumé Les valeurs hématologiques de l'hémoglobine (Hb), l'hématocrite (PCV) et les taux de la folacine (acide folique) et de la cyanocobalamine (vitamine B₁₂) chez des buffles ont été déterminés dans une région de l'Est Haryana, Inde, où sévit le surra à l'état endémique. Les buffles positifs vis-à-vis du surra présentaient des taux significativement faibles de Hb, PCV, d'acide folique et de vitamine B₁₂. Les taux peu élevés de ces deux derniers éléments peuvent renforcer les siqnes cliniques de la trypanosomose chronique, due àT. evansi.

     

Metabolic clustering of a core collection of Indian ginseng Withania somnifera Dunal through DNA, isoenzyme, polypeptide and withanolide profile diversity

Withania somnifera is one of the most important medicinal plants of Ayurveda and finds extensive uses in Indian traditional herbal preparations. In this investigation, selected accessions of the plant were examined for diversity through RAPDs, isoenzymes, polypeptide polymorphism and withanolide profiles. The accessions clustered together with respect to their characteristic profile of major withanolides and represented withaferin A, withanone, withanolide D or withanolide A rich groups. This level of phytochemical diversity as discrete chemotypes is widest and is being first ever documented to occur in Indian population of the plant.     

Isolates of Theileria annulata collected from different parts of India show phenotypic and genetic diversity

Phenotypic and genetic polymorphism was studied amongst four Theileria annulata isolates collected from three different parts of India. Amongst various markers studied for the comparison of growth characteristics of schizont cell lines established from these isolates, viability, non-viability counts and nitric oxide (NO) production showed significant variation. A negative correlation was observed between NO production and mRNA expression for TNF-alpha, a potent proinflammatory cytokine related to the pathogenesis of the disease. Phenotypic polymorphism was also revealed by T. annulata schizont-specific monoclonal antibodies (Mabs), viz. 1C7, 1E11, 2G2 and EU-106, which recognized variable number of cells in indirect fluorescent antibody and indirect immunoperoxidase tests, when tested against the four T. annulata isolates collected from India. Genetic polymorphism was recognized amongst the four isolates by restriction digestion analysis of Tams-1 gene PCR products. These observations revealed that the four isolates of T. annulata are different from each other and might be expressing different antigenic determinants on their cell surface.     

Theileria infectivity ofHyalomma ticks in Haryana,India

Summary Theileria infection of Hyalomma ticks collected from three districts of Haryana was assessed in whole salivary glands by the methyl-green pyronin staining method. Of 1,662 ticks screened, 546 (32·8%) were found Theileria positive. Infection rate in 935 female ticks (36·9%) was more than that of 727 male ticks (27·6%). Density of Theileria infection (number of infected acini per infected tick) was also higher in female ticks.Theileria infection rate of ticks varied greatly in the three districts viz. only 12·0% in Rohtak, 25·8% in Hisar and 48·3% in Karnal. Per cent infection rate was high (63·7%) in ticks from indigenous cattle and low (18·6%) in those collected from buffaloes. However, the intensity of infection in infected ticks from cross-bred cattle was comparatively much higher. Frequency distribution ofTheileria positive acini in infected ticks revealed a low density of infection per infected tick. This points to the largely stable endemic situation prevailing in Haryana. Only a single salivary acinus was found infected in 16·6% of the infected ticks, about 70% had up to 10 infected acini while only about 10% had over 25 infected acini per tick. The masses in acini presumed to beTheileria were confirmed by demonstrating parasitic masses on staining one of a pair of salivary glands and inoculating the suspension of the other half of the gland into two cross-bred cow calves which developed clinical signs and lesions typical of theileriosis.

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Resumen Se determinó la infectividad theilérica de garraptasHyaloma colectadas en tres distritos de Haryana, examinando las glándulas salivares mediante el método de coloración de la pironina verde-metilo. De 1662 ácaros examinados, 546 (32·8%) se encontraron positivas a theileria. La frecuencia de infección en 935 hembras (36·9%) fue más alta que la encontrada en 727 machos (27·6%). El número de acinos infectados por garrapata infectada fue tambien superior en garrapatas hembras. La fracuencia de la infección theilérica vario en tres distritos. Se encontro el 12·0% en Rohtak, 25·8% en Hisar y 48·3% en Karnal. La frequencia de infección por ciento fue alta (63·7%) en garrapatas de ganado criollo y bajo (18·6%) en aquellas colectadas de búfalos. Sin embargo, la frecuencia de infección en garrapatas de ganado cruzado, fue comparativamente más alta. La frecuencia de distribución de acinos positivos a theileria en garrapatas infectadas, reveló una baja densidad de infección por garrapata infectada. Esto se?ala una situación endémica estable prevalente en Haryana. Solamente un acino salivar se encontro infectado en el 16·6% de gappapatas infectadas, cerca del 70·0% tuvo arriba de 10 acinos infectados, mientras que solamente et 10·0% tuvo sobre los 25 acinos infectados, por garrapata. Las masas de acinos presumiblemente de theileria, se confirmaron mediante la tinción de la glándula salivar e inoculando parte de ella en terneros cruzados, los cuales desarrollaron theileriosis.

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Résumé L’infection theilerienne desHyalomma récoltées dans 3 districts de l’Haryana a été examinée sur les glandes salivaires totales par la méthode de coloration au vert de méthyle-pyronine. Sur 1662 tiques, 546 (soit 32,8 p. 100) ont été trouvées positives pour lesTheileria. Le taux d’infection (36,9 p. 100) de 935 tiques femelles est supérieur à celui de 727 males (27,6 p. 100). La densité de l’infection parTheileria (nombre d’acini infectés par tique infectée) est plus grande chez les tiques femelles. Le taux d’infection des tiques varie considérablement selon le district: 12,0 p. 100 dans le Rohtak, 25,8 p. 100 dans l’Hisar et 48,3 p. 100 dans le Karnal. Il est élevé (63,7 p. 100) chez les tiques collectées sur du bétail indigène et bas sur celles prélevées sur des buffles (18,6 p. 100). Cependant, l’intensité de l’infection des tiques prélevées sur du bétail métis est comparativement plus grand. La fréquence de distribution des acini positifs pour lesTheileria chez les tiques infectées révèle une faible intensité d’infection par tique. Ceci montre la situation endémique stable existant dans l’Haryana. On n’a trouvé qu’un seul acinus infecté chez 16,6 p. 100 des tiques infectées; environ 70 p. 100 ont jusqu’à 10 acini infectés tandis que 10 p. 100 ont plus de 25 acini infectés. Les masses présentes dans les acini et présumées être desTheileria ont été confirmées être ces agents par coloration de l’une des paires de glandes salivaires et inoculation de l’autre à deux veaux métis qui ont developpé des signes cliniques et des lésions typiques de theileriose.

     

Evidence for an inter-organismic heme biosynthetic pathway in symbiotic soybean root nodules

The successful symbiosis of soybean with Bradyrhizobium japonicum depends on their complex interactions, culminating in the development and maintenance of root nodules. A B. japonicum mutant defective in heme synthesis in culture was able to produce heme as a result of its symbiotic association with the soybean host. The bacterial mutant was incapable of synthesizing the committed heme precursor delta-aminolevulinic acid (ALA), but nodule plant cells formed ALA from glutamate. In addition, exogenous ALA was taken up by isolated nodule bacteria of the parent strain and of the mutant. It is proposed that bacterial heme found in nodules can be synthesized from plant ALA, hence segments of a single metabolic pathway are spatially separated into two organisms.     

Real-time intraocular pressure measurement during phacoemulsification in dogs ex vivo

This study was performed to evaluate changes in intraocular pressure (IOP) during standard coaxial phacoemulsification using 4 different bottle heights (BHs) and 2 different incision sizes. Coaxial phacoemulsification was performed with a venturi-based machine in 8 enucleated canine eyes through 3.0 and 3.2 mm clear corneal incisions (CCIs). A pressure transducer inserted in the peripheral cornea monitored the IOP in real-time. The surgery was subdivided into 4 stages: sculpt-segment removal, irrigation/aspiration, capsular polishing and viscoelastic removal. The mean IOP and the difference between the maximum and minimum IOPs were calculated at each stage and compared. The ultrasound time and volume of irrigation fluid used were recorded. The mean IOP increased with an elevation in the BH. The mean IOP in the irrigation/aspiration stage was significantly higher than that in the sculpt-segment removal stage at the same BH. The difference between the maximum and minimum IOP at each stage was greater in the 3.2 mm than the 3.0 mm CCIs, although the mean IOP was lower with the 3.2 mm than the 3.0 mm CCIs. The ultrasound time and irrigation fluid volume were greater with the 3.2 mm than the 3.0 mm CCIs. Therefore, fluidic parameters during each stage could be reassessed and adjusted to reduce complications arising from an elevated IOP. Phacoemulsification with 3.0 mm CCIs at a lower BH might lead to less stress on the eye from IOP fluctuations, ultrasound energy and irrigation fluid.     

Seasonal distribution ofTheileria inHyalomma anatolicum anatolicum ticks of an endemic area of haryana,india

Summary The effect of season on the capacity ofHyalomma anatolicum anatolicum ticks to transmitTheileria was studied by detectingTheileria sporoblasts in the salivary glands of 647 adult ticks moulted in winter (November 1990 to March 1991) and 677 adult ticks moulted in the summer-rainy season (June to August 1991). The intensity (number of infected acini per infected tick) and abundance (number of infected acini per tick examined) ofTheileria sporoblasts were significantly (P < 0·05) higher in winter moulted ticks (10·75 and 2·23 respectively) than the summer-rainy season moulted ticks (7·31 and 1·77 respectively). The prevalence of infected moulted ticks was not significantly higher in the summer-rainy season (24·22%) than in the winter (20·71%). A higher percentage of winter moulted ticks had high numbers of infected acini (11/infected tick) than the summer-rainy season moulted ticks, while the trend was reversed for low numbers of infected acini (5/infected tick), being 47·76% for winter and 73·78% for the summer-rainy season. It was concluded that the winter of Haryana is more favourable to the developing theilerial stages in the ticks than the summer-rainy season.

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Distribucion Estacional DeTheileria En La GarrapataHyalomma Anatolicum Anatolicum De Un Area Endemica De Haryana, India

Resumen Se estudió el efecto de la estación del año en la capacidad del la garrapataHyalomma anatolicum anatolicum para transmitirTheileria, detectado esporoblastos deTheileria en las glándulas salivares de 647 garrapatas adultas que mudaron en invierno (noviembre 1990 a marzo 1991) y de 677 garrapatas adultas que mudaron en la estación lluviosa de verano (junio a agosto de 1991). La intensidad (número de acinos infectados por garrapata infectada) y abundancia (número de acinos infectado por garrapata examinada) de esporoblastos deTheileria fue significtavamente (P < 0.05) más alta en las garrapatas que mudaron en invierno (10.75 y 2.23, repectivamente) que en aquellas que mudaron en verano (7.31 y 1.77, respectivamente). La prevaliencia de garrapatas infectadas que mudaron en el verano lluvioso (24.22%), no fue significativamente más alta que en aquellas garrapatas que mudaron en invierno (20·71%). Un porcentaje más alto de garrapatas que mudaron en invierno tuvo el más alto número de acinos infectados (11/garrapatas infectadas) que las garrapatas que mudaron en verano. Lo contrario se encontró para un pequeño número de acinos infectados (5/garrapata infectada), siendo 47.76% para el invierno y 73.78% para el verano lluvioso. Se concluye que el invierno de Haryana es más favorable para el desarrollo de los estadiós deTheileria en las garrapatas que el verano lluvioso.

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Distribution Saisonniere DeTheileria Chez Les TiquesHyalomma Anatolicum Anatolicum D'Une Region Endemique D'Haryana, Inde

Résumé L'influence de la saison sur la capacité des tiquesHyalomma anatolicum anatolicum à transmettréTheileria a été étudié par détection de sporoblastes deTheileria dans les glandes salivaires de 647 tiques adultes écloses en hiver (novembre 1990 à mars 1991) et 677 écloses en saison des pluies d'été (juin à août 1991). L'intensité (nombre d'acini infectés par tique infectée) et l'abondance (nombre d'acini infectés par tique examinée) de sporoblastes deTheileria était significativement plus élevées (P < 0,05) chez les tiques prelevées l'hiver (10,75 et 2,23 respectivement) que chez celles récoltées l'été en saison des pluies (7,31 et 1,77 respectivement). La prévalence des tiques infectées parmi les tiques écloses en saison des pluies d'été (24,22 p.100) n'était pas significativement plus élevé que celle observée parmi les tiques écloses en hiver (20,71 p.100). Un pourcentage élevé de tiques écloses en hiver montrait un nombre plus élevé d'acini infectés (11/tique infectée) que celui des tiques écloses l'été, alors que l'inverse se produisait avec des nombres peu élevés d'acini infectés (5/tique infectée), soit 47,76 p.100 en hiver contre 73,78 p.100 en été. Il en est déduit que l'hiver d'Haryana est plus favorable aux stades de développement des theileries chez les tiques que la saison pluvieuse d'été.